Fluorescence-based discrimination of breast cancer cells by direct exposure to 5-aminolevulinic acid
Fluorescence-based discrimination of breast cancer cells by direct exposure to 5-aminolevulinic acid
复制标题
DOI：
10.1002/cam4.2466
复制
发表时间：
2019-08-05
期刊：
影响因子：
4
通讯作者：
Takamatsu, Tetsuro
中科院分区：
文献类型：
--
作者：
Morita, Midori;Tanaka, Hideo;Takamatsu, Tetsuro
原卟啉IX-荧光测量是口服/局部施用5-氨基乙酰丙酸后用于癌症检测的有力原位方法。然而，这种方法尚未在临床上建立用于乳腺癌，可能是由于5-氨基乙酰丙酸向乳腺的递送不足。在本研究中，我们直接将乳腺癌细胞暴露于5-氨基乙酰丙酸，通过原卟啉IX-荧光来评估其识别能力。通过共聚焦显微镜和流式细胞术测量人乳腺癌细胞系MCF 7和MDA-MB-231以及乳腺上皮细胞系MCF 10A中的荧光强度（FI）。5-氨基乙酰丙酸作用2小时后，MCF 7和MDA-MB-231细胞中的原卟啉IX-FI显著增加，细胞间差异显著，而MCF 10A细胞中的原卟啉IX-FI中度增加。将癌细胞联合暴露于5-氨基乙酰丙酸和ATP结合盒转运蛋白G2的特异性抑制剂Ko 143，进一步增加了原卟啉IX-FI，并减轻了MCF 7和MDA-MB-231细胞中的细胞间变异性，表明基于荧光的癌症检测的重现性和准确性得到改善。在通过细针抽吸从接种MDA-MB-231细胞的小鼠异种移植模型获得的细胞中也证明了这两种药物联合给药的FI增加。此外，截止值增加原卟啉IX-FI比率，之前和之后暴露于这些药物，明确区分癌细胞和非癌细胞。两者合计，直接暴露于5-氨基乙酰丙酸和Ko 143可能是一种有前途的策略，用于使用细针穿刺进行乳腺癌细胞的离体高效荧光检测。
Protoporphyrin IX-fluorescence measurement is a powerful in situ approach for cancer detection after oral/topical administration of 5-aminolevulinic acid. However, this approach has not been clinically established for breast cancer, probably due to insufficient delivery of 5-aminolevulinic acid to the mammary glands. In the present study, we directly exposed breast cancer cells to 5-aminolevulinic acid to assess their discrimination via protoporphyrin IX-fluorescence. Fluorescence intensity (FI) was measured in the human breast cancer cell lines MCF7 and MDA-MB-231 and breast epithelial cell line MCF10A by confocal microscopy and flow cytometry. After 5-aminolevulinic acid exposure for 2 hours, protoporphyrin IX-FI in MCF7 and MDA-MB-231 cells significantly increased with marked cell-to-cell variability, whereas that in MCF10A cells increased moderately. Combined exposure of the cancer cells to 5-aminolevulinic acid and Ko143, a specific inhibitor of ATP-binding cassette transporter G2, further increased protoporphyrin IX-FI and alleviated the cell-to-cell variability in MCF7 and MDA-MB-231 cells, indicating improvement in the reproducibility and accuracy for fluorescence-based cancer detection. The increased FI by combined administration of these two drugs was also demonstrated in cells obtained via fine needle aspiration from mouse xenograft models inoculated with MDA-MB-231 cells. Furthermore, a cutoff value for increased protoporphyrin IX-FI ratio, before and after exposure to these drugs, clearly discriminated between cancer and noncancer cells. Taken together, direct exposure to 5-aminolevulinic acid and Ko143 may be a promising strategy for efficient fluorescence-based detection of breast cancer cells ex vivo using fine needle aspiration.